Effect of DNA methylation of miR-30a-5p promoter region on hepatic injury induced by homocysteine

AIM: To explore the role of DNA methylation of microRNA-30a-5p(miR-30a-5p) promoter region in hepatic injury. METHODS: Four-week-old normal mice and cystathionine β-synthase (CBS) single gene knockout mice were used and divided into normal (CBS^+/+, n=12) group and single gene knockout (CBS^+/-, n=12) group, and the mice were fed with high methionine diet for 8 weeks. HL-7702 hepatic cells were routinely cultured in vitro and divided into control group, homocysteine (Hcy) group and Hcy+5-azacytidne (AZC) group. Serum Hcy, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured by automatic biochemical analyzer. The levels of ALT and AST in the cells culture medium were determined by the microplate method. Hepatic injury in the mice were observed with HE staining. Cell viability staining was used to measure the viability of hepatocytes. RT-qPCR was used to detect the expression of miR-30a-5p in the liver tissues and hepatocytes. The correlation between the expression of miR-30a-5p and serum ALT and AST levels was analyzed by Pearson correlation analysis. DNA methylation level of miR-30a-5p promoter region in the liver tissues and hepatocytes was detected by nested landing methylation-specific PCR (nMS-PCR). RESULTS: Compared with the CBS^+/+ mice, the serum levels of Hcy, ALT and AST in the CBS^+/- mice were significantly increased (P < 0.05). HE staining showed the hepatocyte swelling and nuclear fragmentation and dissolution. The expression level of miR-30a-5p in the liver tissues was decreased (P < 0.01). Besides, the expression level of miR-30a-5p in the mice was negatively correlated with serum ALT and AST levels (r²=0.4557, P=0.0003, r²=0.4626, P=0.0003), and the DNA methylation of miR-30a-5p promoter region was increased (P < 0.01). In the HL-7702 cells, compared with control group,the ALT and AST levels were increased in Hcy group (P < 0.05, P < 0.01), and the cell viability was remarkablely decreased. DNA methylation of miR-30a-5p promoter region was increased (P < 0.01), which decreased after treated the cells with AZC (P < 0.05), while the expression level of miR-30a-5p in the cells was increased (P < 0.05). CONCLUSION: Hypermethylation of miR-30a-5p promoter region may play an important role in hepatic injury.     

番茄匍柄霉叶斑病拮抗细菌的筛选与鉴定

应用平板对峙法从连作多年的番茄根际土样中筛选得到对番茄匍柄霉叶斑病具有较强拮抗活性的细菌菌株ZF161，该菌株对番茄匍柄霉的平板抑制率达到70.51%。离体叶片试验显示，菌株ZF161对番茄匍柄霉叶斑病防治效果达到69.83%。通过菌落形态观察、生理生化特性、Biolog测定和多基因系统发育树综合分析，鉴定菌株ZF161为枯草芽孢杆菌（Bacillus subtilis）。番茄盆栽试验结果表明，菌株ZF161对番茄匍柄霉叶斑病的防治效果达到63.27%。进一步平板对峙抑菌谱试验结果显示，菌株ZF161对其他7种病原真菌也具有较好的抑制效果。上述结果说明，菌株ZF161对番茄匍柄霉叶斑病具有良好的防治效果，且对多种病原真菌也具有抑制作用，具有开发应用的潜力。     

Soluble Klotho protein suppresses THP-1-derived foam cell formation

AIMTo investigate the role of soluble Klotho protein in THP-1-derived foam cell formation. METHODSTHP-1 monocytes were induced into macrophages by treatment with 160 nmol/L phorbol myristate acetate for 48 h, and then were divided into 6 groups: negative control group (THP-1-derived macrophages), positive control group [THP-1-derived foam cells induced by oxidized low-density lipoprotein (ox-LDL) for 48 h], and 25, 50, 100 and 200 μg/L soluble Klotho protein groups (THP-1-derived macrophages pretreated with soluble Klotho protein at the indicat?ed concentraions for 2 h and then induced by ox-LDL for 48 h). Lipid droplets in cytoplasm were observed by oil red O staining. The cholesterol outflow rate was detected by scintillation counting technique. The content of intracellular total cholesterol, free cholesterol and cholesterol ester was detected by enzyme fluorescence analysis. The expression of acyl-coenzyme A:cholesterol acyltransferase 1 （ACAT1） and ATP-binding cassette transport?er A1 (ABCA1) at mRNA and protein levels was determined by RT-qPCR and Western blot, respectively. RESULTSOil red O staining and lipid mass quantification showed that THP-1-derived foam cell formation was dose-dependently suppressed by soluble Klotho protein. The cholesterol efflux rate of THP-1-derived foam cells was increased by soluble Klotho protein in a dose-dependent manner (P<0.05). In addition, soluble Klotho protein decreased the expression of ACAT1 and increased the expression of ABCA1 in a dose-dependent manner (P<0.05). CONCLUSION The soluble Klotho protein inhibits THP-1-derived foam cell formation in a dose-dependent manner by down-regulating the expression of ACAT1 and up-regulating the expression of ABCA1.     

番茄/茄子嫁接对其根际土壤生物学性状及细菌群落结构的影响

以茄子作为砧木，番茄为接穗进行嫁接，并以番茄自根植株和茄子自根植株作为对照，基于传统及现代高通量测序技术分析根际土壤的生物学性状及细菌群落结构特征，旨在揭示嫁接植株抗性增强的机制。结果表明：番茄/茄子嫁接植株根际土壤中可培养细菌、放线菌数量，微生物生物量、酶活性和细菌丰富度、多样性指数均显著高于茄子和番茄自根植株；另一方面，norank_c_Acidobacteria、硝化螺菌属（Nitrospira）、芽孢杆菌属（Bacillus）、norank_f_Gemmatimonadaceae、norank_o_Gaiellales、norank_f_Xanthobacteraceae、norank_o_SC-I-84、norank_f_Anaerolineaceae、norank_f_Nitrosomonadaceae、鞘脂单胞菌属（Sphingomonas）细菌是嫁接植株与茄子、番茄自根植株根际土壤中的共有优势细菌属；g_unclassified_o_Ignavibacteriales 、 醋酸杆菌属（ g_Acetobacter ）、g_norank_f_Longimicrobiaceae 、g_unclassified_f_Verrucomicrobiaceae、g_norank_p_FBP、g_norank_p_Aminicenantes 属细菌是嫁接植株根际土壤的特有优势细菌属。与自根植株相比，番茄/茄子嫁接植株根际土壤存在更高可培养微生物数量，微生物生物量和相关酶活性，具有更高的土壤养分有效性，同时拥有不同占比的共有优势细菌门属，尤其拥有更为丰富的微生物可操作分类单元（operational taxonomic units，OTU）和特有 OTU 数量，这是嫁接植株抗性增强的主要原因。     

羊角椒新品种寿研梦扬的选育

寿研梦扬是由母本CSVHPM1407001和父本CSVHPF1407002配制而成的羊角椒一代杂种。植株长势旺盛,连续坐果能力强;果实长羊角形,果长25～30 cm,果肩宽3～4 cm;青熟果黄绿色,老熟果红色;外表光亮,商品性好,耐贮运;单果质量100 g左右,辣味浓,宜鲜食;田间对病毒病、炭疽病和疫病的抗性强于对照喜洋洋。保护地栽培鲜椒产量可达12 840 kg·(667 m~2)~(-1)左右。适宜山东、河北等地区早春、秋延保护地种植。     

黑甜糯玉米新品种黑甜糯639的选育

黑甜糯639是以糯玉米自交系HNF为母本,以甜玉米自交系D91为父本配制而成的黑甜糯玉米一代杂种。生育期为91 d(天),株高263 cm,穗位151 cm,雄穗主轴与分枝角度中等,一级分枝18～23个,果穗筒型,籽粒黑紫色,甜糯籽粒比例为1∶3,籽粒可溶性糖含量为12.5%,支链淀粉占总淀粉含量的98.1%,口感佳。高抗丝黑穗病、茎腐病、大斑病,抗穗腐病和矮花叶病,平均每667 m~2鲜穗产量900 kg左右。适宜在山西省鲜食糯玉米主产区种植。     

早熟松散型花椰菜新品种优松58的选育

优松58是以细胞质雄性不育系HZ-28为母本,以自交系HL-94为父本配制而成的早熟松散型花椰菜一代杂种。华北地区秋露地从定植至收获60 d(天)左右。株型半直立,生长势强,叶片椭圆形,深绿色,叶缘多波浪;花球半球形,白色,球面光滑,花梗绿,口感脆嫩、甜,品质优,单球质量1.0 kg左右。常规栽培平均每667 m~2产量2 000 kg以上,可适当密植,田间对霜霉病、黑腐病的抗性强于对照津松62,适合北京、天津、河北、河南、山东、安徽等地秋季露地栽培。     

豇豆新品种之豇618的选育

之豇618是以从四川引进的麻子豇豆为母本,以从辽宁引进的ZH优选株系为父本进行杂交,采用系统选育结合分子标记辅助抗枯萎病鉴定育成的中熟豇豆新品种。植株蔓生,花紫色,主侧蔓均可结荚,平均单株结荚14.4条;商品荚油绿色,条荚顺直,喙绿色,鼠尾少,平均荚长63.7 cm,单荚质量26.6 g,豆荚肉厚,粗纤维含量低,口感糯、微甜,可溶性糖含量为24.0 mg · g~(-1);对日照长短不敏感,中抗枯萎病和病毒病,早期产量与之豇106相当,总产量2 000 kg · (667 m~2)~(-1)左右,适宜在全国各地(除高寒地区外)种植。     

辣椒新品种桂牛5号的选育

桂牛5号是以自交系Ca1-5-1为母本,以自交系Ca1-3-8为父本配制而成的牛角椒一代杂种。中早熟,植株生长势较强,连续坐果能力强;果实长牛角形,果长25～30 cm,果肩宽3.5～4.0 cm,单果质量125 g左右;青熟果黄绿色,老熟果鲜红色,商品性好,微辣,果实干物质含量7.88%,VC含量1?530 mg · kg~(-1),辣椒素含量0.023%,宜鲜食,耐贮运;田间对病毒病、炭疽病、疫病的抗性强于对照黄美龙,露地栽培鲜椒产量3?500 kg · (667 m~2)~(-1)左右,适宜广西、广东等地春、秋露地种植。     

叶用芥菜新品种甬雪5号的选育

甬雪5号是以胞质雄性不育系07-50A为母本,以自交系09-3-1为父本配制而成的晚熟叶用芥菜(雪里蕻)一代杂种。播种至采收约112 d(天)。株型开展,生长势强,叶浅绿色,倒卵形,叶缘浅锯齿,叶裂刻深裂,叶面微皱、有光泽。平均有效蘖数26个,单株质量约1.1 kg,一般每667 m~2产量5?000 kg左右。抗病毒病(TuMV),加工品质优良,适宜在长江流域种植。